Method and apparatus for prokaryotic and eukaryotic cell quantitation
AssigneeGenPrime, Inc.
InventorsJames E. Fleming, Jason Buck Somes
Life of this patent
Estimating the term…
Abstract
This invention describes methods and kits for detecting and quantifying viable cells in a sample using fluorescent dyes that can be internalized predominantly by viable cells and have fluorescent properties measurably altered when bound to target components. These methods and kits provide a rapid and cost-effective means of detecting potential biological threats in the field.
Claims22
A method for detecting the presence of viable cells in a sample, comprising:
- a) contacting a sample with a fluorescent dye that binds to target components of a viable cell, wherein said dye is internalized predominately by viable cells and has fluorescence properties that are measurably altered when bound to target components;
- b) detecting total fluorescence of said sample;
- and
- c) comparing the fluorescence detected in step (b) to the fluorescence produced by a control substance, thereby detecting viable cells, wherein the fluorescent dye is SYTO® 16.
A method for detecting the presence of viable cells in a sample, comprising:
- a) contacting a sample with a fluorescent dye that binds to target components of a viable cell, wherein said dye is internalized predominately by viable cells and has fluorescence properties that are measurably altered when bound to target components;
- b) detecting total fluorescence of said sample;
- and
- c) comparing the fluorescence detected in step (b) to the fluorescence produced by a control substance, thereby detecting viable cells, wherein the fluorescent dye binds to DNA of the cells, further comprising treating the sample with DNase before contacting the sample with the fluorescent dye.
A method for detecting the presence of viable cells in a sample, comprising:
- a) contacting a sample with a fluorescent dye that binds to target components of a viable cell, wherein said dye is internalized predominately by viable cells and has fluorescence properties that are measurably altered when bound to target components;
- b) detecting total fluorescence of said sample;
- and
- c) comparing the fluorescence detected in step (b) to the fluorescence produced by a control substance, thereby detecting viable cells, wherein the fluorescent dye is SYTO® 16, further comprising correlating the fluorescence detected in step (b) to the number of viable cells in the sample.
A method for detecting the presence of viable cells in a sample, comprising:
- a) contacting a sample with a fluorescent dye that binds to target components of a viable cell, wherein said dye is internalized predominately by viable cells and has fluorescence properties that are measurably altered when bound to target components;
- b) detecting total fluorescence of said sample;
- and
- c) comparing the fluorescence detected in step (b) to the fluorescence produced by a control substance, thereby detecting viable cells, wherein the fluorescent dye binds to DNA of the cells, further comprising treating the sample with DNase before contacting the sample with the fluorescent dye, further comprising correlating the fluorescence detected in step (b) to the number of viable cells in the sample.
A kit for detecting viable cells in a sample, comprising a cell suspension solution, a fluorescent dye that can be internalized predominantly by viable cells, and instructions for detecting dye binding to cellular components of viable cells.
A kit for detecting or quantifying viable cells, comprising:
- instructions for detecting or quantifying viable cells, a first container containing a first solution, means for placement of a sample containing an unknown number of viable cells into a solution containing a fluorescent dye that can be internalized predominately by the viable cells and binds selectively to double-stranded DNA or other specific cellular components, whereupon its fluorescence is altered to a measurable degree, and means for illuminating the mixture of first solution with said sample with excitation light and measuring fluorescence emitted by said mixture, thereby detecting the presence of viable cells in said sample.
A kit for detecting or quantifying viable cells, comprising:
- instructions for detecting or quantifying viable cells, a first container containing a first solution, means for placement of a sample containing an unknown number of viable cells into a first solution, means for concentrating the solids and cells from the mixture of said first solution with said sample and retaining said solids from the remainder of said mixture, a second solution containing a fluorescent dye that can be internalized predominately by the viable cells and binds selectively to double-stranded DNA or other specific cellular components, whereupon its fluorescence is altered to a measurable degree, means for mixing said second solution with said solids to form a second mixture, and means for illuminating the mixture of said second solution with said solids with excitation light and measuring fluorescence emitted by said mixture, thereby detecting viable cells in said sample.
Dependencies are inferred from the claim text.